Supplementary material

Figure S1: Sizes of predicted amplicons according to primer pair. Colors indicate the threshold for the number of accepted mismatches in the in silico PCR run. Off-target amplicons were removed from this figure.

Figure S2: Virtual gel depicting the smallest and largest in silico predicted target amplicon for each primer pair (maximum number of allowed mismatches: 3). Image was created with the Virtual Gel function in Geneious (Biomatters) using the MW Ladder: 100 bp (NEB).

File S1: Reduced version of the alignment originally published in Dubuffet et al. [9], only including sequences that we selected for the in silico analyses.

File S2: Visualization of the reduced alignment originally published in Dubuffet et al. [9] showing the primer bindings. The image was created in Geneious (Biomatters) with the maximum of allowed mismatches set to three. Note that some off-targets appear due to longer sequence regions consisting of “N”s which did not appear in the actual analyses and were therefore interpreted as erroneous.

Table S1: Selection of example literature in which the analyzed primer pairs were used.

Table S2: Overview of the analyzed sequences that were selected from an alignment published in Dubuffet et al. [9].

Table S3: Number of amplified species within each clade, according to primer pair and number of allowed mismatches in the corresponding in silico PCR run. Off-targets are excluded to avoid multiple counting of the same microsporidian.

Table S4: Amplicon sizes according to primer pair and number of accepted mismatches in the corresponding in silico PCR run.

Table S5: Identified mismatch combinations for each individual primer tested.

Table S6: Overview of off-target amplicons identified in the in silico PCR runs for primer pairs only.

Table S7: Overview of off-target bindings identified in the in silico PCR runs for each individual primer.

© A. Doliwa et al., published by EDP Sciences, 2023